Oct 25, 2020
DNT Induction
- 12020 iGEM NEFU China
- 2020 iGEM NEFU China
Protocol Citation: Zhujun Wei 2020. DNT Induction. protocols.io https://dx.doi.org/10.17504/protocols.io.bnxsmfne
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 25, 2020
Last Modified: October 25, 2020
Protocol Integer ID: 43730
The DNT powder (>99%) was dissolved in 80% acetonitrile solution,prepared as 10g/L stock solution, and stored at 4℃.
The glycerin strains were first grown overnight in LB medium at 37℃supplied with appropriate antibiotics.
The bacteria cultured overnight were diluted by 1/100 times into fresh LB medium, and a certain amount of DNT stock was added to make DNT reach an expected concentration. Then, the samples (200 µL) were added to the wells of a clear 96-well platewith LB medium only as a negative control.
The RFU(relative fluorescence units) and OD600 were measured every 30 minutes using the multi-functional enzyme readerat 37°C shaking culture condition, for a period of 12h.(For RFU measurement, 200 µL of LB was used for calibration, with an excitation wavelength 485nm, an excitation bandwidth 20nm, an emission wavelength 535nm, and an emission width 70nm)
The experiment was repeated three times for each sample.